IL43343A - Production of plasminogen activator - Google Patents
Production of plasminogen activatorInfo
- Publication number
- IL43343A IL43343A IL43343A IL4334373A IL43343A IL 43343 A IL43343 A IL 43343A IL 43343 A IL43343 A IL 43343A IL 4334373 A IL4334373 A IL 4334373A IL 43343 A IL43343 A IL 43343A
- Authority
- IL
- Israel
- Prior art keywords
- plasminogen activator
- mammalian cells
- medium
- atcc
- buffer
- Prior art date
Links
- 102000001938 Plasminogen Activators Human genes 0.000 title claims 24
- 108010001014 Plasminogen Activators Proteins 0.000 title claims 24
- 229940127126 plasminogen activator Drugs 0.000 title claims 24
- 238000000034 method Methods 0.000 claims 19
- 210000004962 mammalian cell Anatomy 0.000 claims 10
- 210000003292 kidney cell Anatomy 0.000 claims 7
- 239000002609 medium Substances 0.000 claims 7
- 150000003839 salts Chemical class 0.000 claims 6
- 229930013930 alkaloid Natural products 0.000 claims 5
- 150000003797 alkaloid derivatives Chemical class 0.000 claims 5
- IAKHMKGGTNLKSZ-INIZCTEOSA-N (S)-colchicine Chemical compound C1([C@@H](NC(C)=O)CC2)=CC(=O)C(OC)=CC=C1C1=C2C=C(OC)C(OC)=C1OC IAKHMKGGTNLKSZ-INIZCTEOSA-N 0.000 claims 4
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims 4
- 239000000872 buffer Substances 0.000 claims 4
- 229910052588 hydroxylapatite Inorganic materials 0.000 claims 4
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 claims 4
- 239000003104 tissue culture media Substances 0.000 claims 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims 4
- 229910017053 inorganic salt Inorganic materials 0.000 claims 3
- 235000015097 nutrients Nutrition 0.000 claims 3
- 239000008363 phosphate buffer Substances 0.000 claims 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims 2
- 241000282560 Macaca mulatta Species 0.000 claims 2
- JXLYSJRDGCGARV-WWYNWVTFSA-N Vinblastine Natural products O=C(O[C@H]1[C@](O)(C(=O)OC)[C@@H]2N(C)c3c(cc(c(OC)c3)[C@]3(C(=O)OC)c4[nH]c5c(c4CCN4C[C@](O)(CC)C[C@H](C3)C4)cccc5)[C@@]32[C@H]2[C@@]1(CC)C=CCN2CC3)C JXLYSJRDGCGARV-WWYNWVTFSA-N 0.000 claims 2
- 239000003463 adsorbent Substances 0.000 claims 2
- 239000007853 buffer solution Substances 0.000 claims 2
- 238000004113 cell culture Methods 0.000 claims 2
- -1 colchamide Chemical compound 0.000 claims 2
- 229960001338 colchicine Drugs 0.000 claims 2
- 238000012258 culturing Methods 0.000 claims 2
- 239000001103 potassium chloride Substances 0.000 claims 2
- 235000011164 potassium chloride Nutrition 0.000 claims 2
- 239000007787 solid Substances 0.000 claims 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims 2
- 229960003048 vinblastine Drugs 0.000 claims 2
- JXLYSJRDGCGARV-XQKSVPLYSA-N vincaleukoblastine Chemical compound C([C@@H](C[C@]1(C(=O)OC)C=2C(=CC3=C([C@]45[C@H]([C@@]([C@H](OC(C)=O)[C@]6(CC)C=CCN([C@H]56)CC4)(O)C(=O)OC)N3C)C=2)OC)C[C@@](C2)(O)CC)N2CCC2=C1NC1=CC=CC=C21 JXLYSJRDGCGARV-XQKSVPLYSA-N 0.000 claims 2
- 229960004528 vincristine Drugs 0.000 claims 2
- OGWKCGZFUXNPDA-XQKSVPLYSA-N vincristine Chemical compound C([N@]1C[C@@H](C[C@]2(C(=O)OC)C=3C(=CC4=C([C@]56[C@H]([C@@]([C@H](OC(C)=O)[C@]7(CC)C=CCN([C@H]67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)C[C@@](C1)(O)CC)CC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-XQKSVPLYSA-N 0.000 claims 2
- OGWKCGZFUXNPDA-UHFFFAOYSA-N vincristine Natural products C1C(CC)(O)CC(CC2(C(=O)OC)C=3C(=CC4=C(C56C(C(C(OC(C)=O)C7(CC)C=CCN(C67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)CN1CCC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-UHFFFAOYSA-N 0.000 claims 2
- IPDJRYUVROTJTO-UHFFFAOYSA-N 1,2,3,10-tetramethoxy-6,7-dihydro-5h-benzo[a]heptalen-9-one Chemical compound C1CCC2=CC(=O)C(OC)=CC=C2C2=C1C=C(OC)C(OC)=C2OC IPDJRYUVROTJTO-UHFFFAOYSA-N 0.000 claims 1
- JVIPLYCGEZUBIO-UHFFFAOYSA-N 2-(4-fluorophenyl)-1,3-dioxoisoindole-5-carboxylic acid Chemical compound O=C1C2=CC(C(=O)O)=CC=C2C(=O)N1C1=CC=C(F)C=C1 JVIPLYCGEZUBIO-UHFFFAOYSA-N 0.000 claims 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims 1
- HFPMXDMZJUJZBX-AWEZNQCLSA-N Deacetylcolchicine Chemical compound C1([C@@H](N)CC2)=CC(=O)C(OC)=CC=C1C1=C2C=C(OC)C(OC)=C1OC HFPMXDMZJUJZBX-AWEZNQCLSA-N 0.000 claims 1
- 229920001425 Diethylaminoethyl cellulose Polymers 0.000 claims 1
- 241000283973 Oryctolagus cuniculus Species 0.000 claims 1
- 229920005654 Sephadex Polymers 0.000 claims 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical group N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims 1
- 235000011130 ammonium sulphate Nutrition 0.000 claims 1
- 229910052799 carbon Inorganic materials 0.000 claims 1
- 210000004027 cell Anatomy 0.000 claims 1
- 239000006285 cell suspension Substances 0.000 claims 1
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 238000007865 diluting Methods 0.000 claims 1
- 239000012153 distilled water Substances 0.000 claims 1
- 239000012535 impurity Substances 0.000 claims 1
- 229910052757 nitrogen Inorganic materials 0.000 claims 1
- 239000002244 precipitate Substances 0.000 claims 1
- 238000002360 preparation method Methods 0.000 claims 1
- 239000000243 solution Substances 0.000 claims 1
- 238000005406 washing Methods 0.000 claims 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
- C12N9/6424—Serine endopeptidases (3.4.21)
- C12N9/6456—Plasminogen activators
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Claims (19)
1. Λ process for the preparation of plasminogen activator which comprises : a) culturing plasminogen activator producing mammalian cells in an aqueous nutrient tissue culture medium containing assimilable sources of nitrogen, carbon and inorganic salts to obtain a viable cell culture; b) culturing said viable cell culture in an aqueous nutrient tissue culture medium containing between 0.1 and 10 mcg/ml . of an alkaloid selected from the group consisting of colchicine, colchamide, desacetylcolchicine , desacetamidocolchicine , cyanocolchicine , vinblastine and vincristine until a substantial amount of plasminogen activator is present in said medium; and c) recovering the plasminogen activator from said tissue culture medium.
2. The process of claim 1 wherein the mammalian cells are cultured in the presence of an inert solid support to form a viable culture of confluent cells on said support.
3. The process of claim 1 wherein the mammalian cells are cultured to maximum population density in a viable, submerged cell suspension 'culture.
4. The process of any of claims 1, 2 or 3 wherein the mammalian cells are mammalian kidney cells.
5. The process of any of claims 1-4 in which the mammalian cells are porcine kidney cells PK (15) ; ATCC No. CCL 33 PK (15) . 0
6. The process of any of claims 1-4 in which the mammalian cells are porcine kidney cells LLC-PK, (Hull), ATCC No. CL 101.
7. The process of any of claims 1-4 in which the mammalian cells are rabbit kidney cells (Hull) , ATCC No. CCL 106.
8. The process of any of claims 1-4 in which the mammalian cells are rhesus monkey kidney cells LLC-MI<2 (Hull) , ATCC No. CCL 7.
9. The process of any of claims 1-4 in which the mammalian cells are rhesus monkey kidney cells LLC-MI^/ ATCC No. CCL 7.1.
10. The process of any of claims 1-4 in which the mammalian cells are porcine kidney cells LLC-PK^, ATCC No. CL 101.1.
11. The process of any of claims 1-10 wherein the alkaloid is colchicine.
12. The process of any of claims 1-10 wherein the alkaloid is colchamide.
13. The process of any of claims 1-10 wherein the alkaloid is vinblastine.
14. The process of any of claims 1-10 wherein the alkaloid is vincristine.
15. The process of claim 1 wherein a purified plasminogen activator is recovered from an aqueous nutrient tissue culture medium by. a) separating insolubles from the aqueous tissue medium containing plasminogen activator ; b) diluting the separated medium with distilled water to adjust the specific conductance of said medium to 8 millir.hos or less; c) contacting said plasminogen activator containing medium for between 1 and 24 hours with between 3 and 12 g. of hydroxyapatite per liter of medium; d) separating the hydroxyapatite containing adsorbed plasminogen activator from said medium; e) washing said hydroxyapatite; f) eluting said plasminogen activator from the washed hydroxyapatite with 0.7 M phosphate buffer at pH 6.8; g) adding to the plasminogen activator containing eluate a water soluble inorganic salt in an amount sufficient to achieve a salt concentration of appro imately 20 percent of saturation ; h) separating the precipitated impurities from said eluate; i) adding to said eluate an additional amount of said salt to achieve a salt concentration of between approximately 45 and 90 percent of saturation; j) separating the precipitated plasminogen activator from said salt containing eluate; k) dissolving the plasminogen activator in phosphate buffer having a molarity of between approximately 0.001 and 0.005 and dialyzing said buffer solution; 1) contacting the dialyzed solution of plasminogen activator with diethylaminoethyl cellulose adsorbent; m) eluting the adsorbed plasminogen activator from said adsorbent with 0.005 molar phosphate buffer; n) adding to the plasminogen activator containing eluate a water soluble inorganic salt in an amount sufficient to achieve a salt concentration of approximately 75 percent of saturation; o) separating the precipitate of plasminogen activator; 10 p) dissolving the plasminogen activator in 0.1 tris-hydroxymethylaminomethane : 0.1 M potassium chloride buffer, pll 8.0 and dialyzing the buffer against said buffer alone; q) contacting the dialyzed buffer solution with a modified dextran gel; r) eluting the adsorbed plasminogen activator from said gel with 0.1 M tris-hydroxymethylamino- methane:0.1 14 potassium chloride, pH 8.0 buffer; and 20 s) lyophilizing the plasminogen activator containing eluate to obtain in a purified solid form the plasminogen activator.
16. The method of claim 15, in which the water soluble inorganic salt is ammonium sulfate.
17. The process of preparing plasminogen activator substantially as herein described with particular reference to any one of the specific examples.
18. The process of preparing plasminogen activator substantially as herein described with particular reference 30 to any of specific Examples 1-4.
19. The process of preparing plasminogen activator substantially as herein described v/ith particular reference to Example 5. S. HOROWITZ & CO. AGENTS FOR APPLICANTS
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US30005472A | 1972-10-24 | 1972-10-24 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| IL43343A0 IL43343A0 (en) | 1973-11-28 |
| IL43343A true IL43343A (en) | 1976-03-31 |
Family
ID=23157506
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| IL43343A IL43343A (en) | 1972-10-24 | 1973-09-30 | Production of plasminogen activator |
Country Status (9)
| Country | Link |
|---|---|
| JP (1) | JPS4975794A (en) |
| BE (1) | BE806323A (en) |
| CH (1) | CH589142A5 (en) |
| DE (1) | DE2353318A1 (en) |
| FR (1) | FR2203876B1 (en) |
| GB (1) | GB1443189A (en) |
| IL (1) | IL43343A (en) |
| NL (1) | NL7314579A (en) |
| SE (2) | SE392618B (en) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5571496A (en) * | 1978-11-24 | 1980-05-29 | Asahi Chem Ind Co Ltd | Preparation of living substance |
| US5185259A (en) * | 1982-05-05 | 1993-02-09 | Genentech, Inc. | Truncated human tissue plasminogen activator |
| FI831484L (en) * | 1982-05-05 | 1983-11-06 | Genentech Inc | PLASMINOGEN AKTIVATOR FOER MAENSKOVAEVNAD |
| SU1662352A3 (en) * | 1982-05-05 | 1991-07-07 | Генентек, Инк (Фирма) | Strain of bacteria escherichia coli - a producer of plasminogen activator of tissue type |
| IL68561A (en) * | 1982-05-05 | 1991-01-31 | Genentech Inc | Preparation of polypeptide with human tissue plasminogen activator function,processes for making it,and dna and transformed cell intermediates thereof |
| US4853330A (en) * | 1983-04-07 | 1989-08-01 | Genentech, Inc. | Human tissue plasminogen activator |
| IL69664A0 (en) * | 1982-09-15 | 1983-12-30 | Collaborative Res Inc | Medium supplement and method for growing cells in vitro |
| HU199883B (en) * | 1984-09-21 | 1990-03-28 | Boehringer Ingelheim Int | Process for producing anticoagulant proteins and pharmaceutical compositions comprising same |
| JP2507339B2 (en) * | 1986-08-11 | 1996-06-12 | 三井東圧化学株式会社 | Purification method of crude tPA |
-
1973
- 1973-09-30 IL IL43343A patent/IL43343A/en unknown
- 1973-10-19 FR FR7337349A patent/FR2203876B1/fr not_active Expired
- 1973-10-22 BE BE1005443A patent/BE806323A/en unknown
- 1973-10-22 CH CH1484773A patent/CH589142A5/xx not_active IP Right Cessation
- 1973-10-23 GB GB4919673A patent/GB1443189A/en not_active Expired
- 1973-10-23 SE SE7314381A patent/SE392618B/en unknown
- 1973-10-23 NL NL7314579A patent/NL7314579A/xx not_active Application Discontinuation
- 1973-10-24 DE DE19732353318 patent/DE2353318A1/en active Pending
- 1973-10-24 JP JP48119815A patent/JPS4975794A/ja active Pending
-
1976
- 1976-09-07 SE SE7609880A patent/SE7609880L/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| SE7609880L (en) | 1976-09-07 |
| IL43343A0 (en) | 1973-11-28 |
| FR2203876B1 (en) | 1977-05-27 |
| GB1443189A (en) | 1976-07-21 |
| NL7314579A (en) | 1974-04-26 |
| SE392618B (en) | 1977-04-04 |
| FR2203876A1 (en) | 1974-05-17 |
| JPS4975794A (en) | 1974-07-22 |
| CH589142A5 (en) | 1977-06-30 |
| DE2353318A1 (en) | 1974-05-02 |
| BE806323A (en) | 1974-04-22 |
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